[Histonet] Re: Hydrogen Peroxide quenching

[Ford, Judi]

Okay, so I'm confused. Some labs do the hydrogen peroxide step before HIER and others afterwards, why? Also, are there differences using hydrogen peroxide diluted in water/methanol/or PBS? Some bring paraffin slides down to absolute alcohol then do the hydrogen peroxide step where others wait until bringing the slides to water before doing the step. Is there some rule of thumb, or do you just figure out which procedure works for that specific antibody in your lab?

I'm still pretty new to this, so sorry if this seems like a dumb question.

Judi F.
Palo Alto, CA

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Wednesday, September 26, 2007 2:49 PM
To: Daivik_Shah <@t> rush.edu; histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Re: Hydrogen Peroxide quenching

You use it immediately after the HIER, before starting the IHC protocol.
  René J.

Daivik_Shah <@t> rush.edu wrote:
  Hello friends,
I have question regarding Hydrogen peroxide quenching. Is this treatment 
applied in Immunohistostain after deparafinization or before application 
of secondary antibody? Is that affect any way to stain?

Thanks.
Daivik
_____________________________
Daivik Shah, Histotechnologist, M.S. Biotechnology
RADC Laboratory, Cohn Building 436
Rush University Medical Center
Tel. 312-563-4891


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